L-Carnitine Regulates The Proliferation And Migration Of Prostate Cancer Through MBD2-HDAC1/2-hepaCAM

Objective:To investigate the effect of proliferation and migration of prostate cancer(PCa)under the supplementation of L-carnitine(LC)and its molecular mechanism.Methods:(1)The database was used to predict the expression of MBD2 and HDAC1/2 in PCa tissues and adjacent tissues,and the interaction and correlation between MBD2 and HDAC1/2 were predicted by bioinformatics method.Collected the PCa tissue and adjacent nonmalignant tissue,the protein expression of MBD2 、 HDAC1/2 and hepaCAM were detected by immunohistochemistry(IHC)and their correlation were verified by Spearman analysis.(2)Q-PCR and WB to detect the expression of MBD2 、 HDAC1 、 HDAC2 、 Ac-H3K9 and hepaCAM after treated with sh-MBD2 、 HDAC inhibitor and adenovirus-hepaCAM adenovirus(Ad-hepCAM).(3)The effect of LC on proliferation of PCa cells was determined by MTT assay.PCa cells were treated with different concentrations of LC(0、4、8mM)for 72 h.Then transwell and wound healing assay were used to detect the migration;colony formation assay to detect the colony formation ability;Q-PCR and WB to detect the expression of MBD2、HDAC1、HDAC2、Ac-H3K9 and hepaCAM.(4)Tumor xenografts were established by injecting PC3 cells subcutaneously to nude mice.LC was injected intraperitoneally,the tumor volume was measured every 3 days,after 15 days,Q-PCR and IHC to detect the expression of MBD2 、 HDAC1 、HDAC2 and hepaCAM.Results:(1)The database predicted that the expression of MBD2 and HDAC1/2 were different in PCa tissues and adjacent tissues,and the bioinformatics method predicted that MBD2 and HDAC1/2 had a close interaction and significant positive correlation.IHC results showed that the expression of MBD2 、 HDAC1 and HDAC2 of PCa tissue were higher significantly than normal control,while the expression of hepaCAM was reversed(p<0.05);Spearman analysis suggested that their expression were negatively correlated(MBD2:r=-0.680,p<0.05;HDAC1:r=-0.645,p<0.05;HDAC2:r=-0.641,p<0.05).(2)The expression of MBD2 、 HDAC1 and HDAC2 were significantly decreased,Ac-H3K9 and hepaCAM were increased after being treated with sh-MBD2(p<0.05);The expression of MBD2、HDAC1 and HDAC2 were significantly decreased,Ac-H3K9 and hepaCAM were increased after being treated with HDAC inhibitor(p<0.05);There were no significant differences in the expression of MBD2、HDAC1 and HDAC2 in adenovirus-hepaCAM adenovirus group and adenovirus-GFP adenovirus group(p>0.05).(3)LC displayed a growth-inhibiting effect on PCa cells in a dose and time-dependent manner.After treating PCa cells with different concentrations of LC(0、4、8mM)for72h,the cell migration capacity were significantly restrained,the colony formation ability were obviously reduced(p<0.05);the expression of MBD2、HDAC1 and HDAC2 were significantly decreased,Ac-H3K9 and hepaCAM were increased(p<0.05).(4)After LC treatment,the volume oftumor xenografts were remarkably reduced,the expression of MBD2 、HDAC1 and HDAC2 were significantly decreased and the expression of hepaCAM was increased(p<0.05).Conclusions:LC may inhibit the proliferation and migration of PCa cells via MBD2-HDAC1/2 and reverse the expression of hepaCAM.